Correlative scanning electron and confocal microscopy imaging of labeled cells coated by indium-tin-oxide

Handle URI:
http://hdl.handle.net/10754/564109
Title:
Correlative scanning electron and confocal microscopy imaging of labeled cells coated by indium-tin-oxide
Authors:
Rodighiero, Simona; Torre, Bruno; Sogne, Elisa ( 0000-0003-2097-4358 ) ; Ruffilli, Roberta; Cagnoli, Cinzia; Francolini, Maura; Di Fabrizio, Enzo M. ( 0000-0001-5886-4678 ) ; Falqui, Andrea ( 0000-0002-1476-7742 )
Abstract:
Confocal microscopy imaging of cells allows to visualize the presence of specific antigens by using fluorescent tags or fluorescent proteins, with resolution of few hundreds of nanometers, providing their localization in a large field-of-view and the understanding of their cellular function. Conversely, in scanning electron microscopy (SEM), the surface morphology of cells is imaged down to nanometer scale using secondary electrons. Combining both imaging techniques have brought to the correlative light and electron microscopy, contributing to investigate the existing relationships between biological surface structures and functions. Furthermore, in SEM, backscattered electrons (BSE) can image local compositional differences, like those due to nanosized gold particles labeling cellular surface antigens. To perform SEM imaging of cells, they could be grown on conducting substrates, but obtaining images of limited quality. Alternatively, they could be rendered electrically conductive, coating them with a thin metal layer. However, when BSE are collected to detect gold-labeled surface antigens, heavy metals cannot be used as coating material, as they would mask the BSE signal produced by the markers. Cell surface could be then coated with a thin layer of chromium, but this results in a loss of conductivity due to the fast chromium oxidation, if the samples come in contact with air. In order to overcome these major limitations, a thin layer of indium-tin-oxide was deposited by ion-sputtering on gold-decorated HeLa cells and neurons. Indium-tin-oxide was able to provide stable electrical conductivity and preservation of the BSE signal coming from the gold-conjugated markers. © 2015 Wiley Periodicals, Inc.
KAUST Department:
Physical Sciences and Engineering (PSE) Division; Biological and Environmental Sciences and Engineering (BESE) Division; Materials Science and Engineering Program; Bioscience Program
Publisher:
Wiley-Blackwell
Journal:
Microscopy Research and Technique
Issue Date:
22-Mar-2015
DOI:
10.1002/jemt.22492
Type:
Article
ISSN:
1059910X
Appears in Collections:
Articles; Bioscience Program; Physical Sciences and Engineering (PSE) Division; Materials Science and Engineering Program; Biological and Environmental Sciences and Engineering (BESE) Division

Full metadata record

DC FieldValue Language
dc.contributor.authorRodighiero, Simonaen
dc.contributor.authorTorre, Brunoen
dc.contributor.authorSogne, Elisaen
dc.contributor.authorRuffilli, Robertaen
dc.contributor.authorCagnoli, Cinziaen
dc.contributor.authorFrancolini, Mauraen
dc.contributor.authorDi Fabrizio, Enzo M.en
dc.contributor.authorFalqui, Andreaen
dc.date.accessioned2015-08-03T12:32:31Zen
dc.date.available2015-08-03T12:32:31Zen
dc.date.issued2015-03-22en
dc.identifier.issn1059910Xen
dc.identifier.doi10.1002/jemt.22492en
dc.identifier.urihttp://hdl.handle.net/10754/564109en
dc.description.abstractConfocal microscopy imaging of cells allows to visualize the presence of specific antigens by using fluorescent tags or fluorescent proteins, with resolution of few hundreds of nanometers, providing their localization in a large field-of-view and the understanding of their cellular function. Conversely, in scanning electron microscopy (SEM), the surface morphology of cells is imaged down to nanometer scale using secondary electrons. Combining both imaging techniques have brought to the correlative light and electron microscopy, contributing to investigate the existing relationships between biological surface structures and functions. Furthermore, in SEM, backscattered electrons (BSE) can image local compositional differences, like those due to nanosized gold particles labeling cellular surface antigens. To perform SEM imaging of cells, they could be grown on conducting substrates, but obtaining images of limited quality. Alternatively, they could be rendered electrically conductive, coating them with a thin metal layer. However, when BSE are collected to detect gold-labeled surface antigens, heavy metals cannot be used as coating material, as they would mask the BSE signal produced by the markers. Cell surface could be then coated with a thin layer of chromium, but this results in a loss of conductivity due to the fast chromium oxidation, if the samples come in contact with air. In order to overcome these major limitations, a thin layer of indium-tin-oxide was deposited by ion-sputtering on gold-decorated HeLa cells and neurons. Indium-tin-oxide was able to provide stable electrical conductivity and preservation of the BSE signal coming from the gold-conjugated markers. © 2015 Wiley Periodicals, Inc.en
dc.publisherWiley-Blackwellen
dc.subjectConfocal microscopyen
dc.subjectCorrelative microscopyen
dc.subjectImmunolabelingen
dc.subjectITOen
dc.subjectScanning electron microscopyen
dc.titleCorrelative scanning electron and confocal microscopy imaging of labeled cells coated by indium-tin-oxideen
dc.typeArticleen
dc.contributor.departmentPhysical Sciences and Engineering (PSE) Divisionen
dc.contributor.departmentBiological and Environmental Sciences and Engineering (BESE) Divisionen
dc.contributor.departmentMaterials Science and Engineering Programen
dc.contributor.departmentBioscience Programen
dc.identifier.journalMicroscopy Research and Techniqueen
dc.contributor.institutionFondazione Filarete, Viale Ortles 22/4, Milano, Italyen
dc.contributor.institutionCEMES/CNRS, 29 Rue Jeanne Marvig BP 94347, Toulouse Cedex 4, Franceen
dc.contributor.institutionDepartment of Medical Biotechnology and Translational Medicine, Università Degli Studi Di Milano, Milano, Italyen
kaust.authorTorre, Brunoen
kaust.authorSogne, Elisaen
kaust.authorDi Fabrizio, Enzo M.en
kaust.authorFalqui, Andreaen
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